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Comment: Updated Symphony Baseline Report

Table of Contents
maxLevel2

...

Laser (nm)

Power (mw)DetectorCommon fluorochromes detectable (not exhaustive)Band pass filterDichroic MirrorLaser orderrSDen2.5x rSDen

405

50





1



V450_50B

Pacific Blue, Cascade Blue, AF405, V450, BV421, DAPI, eFluor450, SparkViolet423, SuperBright436

450/50

NA


23.5

58.75



V510_50A

CFP, AF430, AmCyan, Cascade Yellow, V500, BV510, PO,eFluor506, SparkViolet538

510/50

502LP


22

55

488 

20





2



SSC

SSC

488/10

NA


NA

 NA



B530_30E

FITC, GFP, AF488, YFP, 5-FAM, CFSE, BB515, ZsGreen, SYBR Green, BODIPY FL

530/30

502LP


22.1

55.25



B585_42D

PE,PI,DsRed,DiA

585/42

556LP


19.8

49.5



B670_LPB

PerCP,PI, PerCP-Cy5.5,PerCP-Cy5,PE-Cy5,7AAD, BB700

670LP

655LP


21.7

54.25



B780_60A

PE-Cy7, PE-Vio770

780/60

735LP


21.8

54.8

633

20





3



R660_20C

APC, AF647, CF647, DiD, DRAQ5, eFluor660

660/20

NA


21.7

54.25



R780_60A

APC-Cy7, APC-H7, DiR, APC-FIRE, APC-eFluor780, APCVio770

780/60

735LP


21.8

54.5

...

Laser (nm)Power (mw)DetectorCommon fluorochromes detectable (not exhaustive)Band pass filterDichroic MirrorLaser orderrSDen2.5x rSDen

405

50





1



V450_50B

Pacific Blue, Cascade Blue, AF405, V450, BV421, DAPI, eFluor450, SparkViolet423, SuperBright436

450/50

NA


1640



V510_50A

CFP, AF430, AmCyan, Cascade Yellow, V500, BV510, PO, eFluor506, SparkViolet538

510/50

502LP


16.140.25

488 

20





2



SSC

SSC

488/10

NA


N/A


B530_30E

FITC, GFP, AF488, YFP, 5-FAM, CFSE, BB515, ZsGreen, SYBR Green, BODIPY FL

530/30

502LP

 

21.353.25


B585_42D

PE,PI,DsRed,DiA

585/42

556LP


22.957.25


B670_LPB

PerCP,PI, PerCP-Cy5.5,PerCP-Cy5,PE-Cy5,7AAD, BB700

670LP

655LP


22.255.5


B780_60APE-Cy7, PE-Vio770

780/60

735LP


21.152.75

633

20





3



R660_20C

APC, AF647, CF647, DiD, DRAQ5, eFluor660

660/20

NA


16.140.25


R780_60A

APC-Cy7, APC-H7, DiR, APC-FIRE, APC-eFluor780, APCVio770

780/60

735LP


15.639

...

Laser (nm)Power (mw)DetectorCommon fluorochromes detectable (not exhaustive)Band pass filterDichroic MirrorLaser orderrSDen2.5x rSDen

488 

20





1



SSC

SSC

488/10

NA


N/A


Alexa Fluor 488

FITC, GFP, AF488, YFP, 5-FAM, CFSE, BB515, ZsGreen, SYBR Green, BODIPY FL

530/30

502LP

 

22.857


PE

PE,PI,DsRed,DiA

585/42

556LP


22.857


PerCP

PerCP,PI, PerCP-Cy5.5,PerCP-Cy5,PE-Cy5,7AAD, BB700

670LP

655LP


22.857


PE-Cy7PE-Cy7, PE-Vio770

780/60

735LP


20.551.25

633

20





2



APC

APC, AF647, CF647, DiD, DRAQ5, eFluor660

660/20

NA


22.857


APC-Cy7

APC-Cy7, APC-H7, DiR, APC-FIRE, APC-eFluor780, APCVio770

780/60

735LP


23.358.25

...

Laser (nm)Power (mw)DetectorCommon fluorochromes detectable (not exhaustive)Band pass filterDichroic MirrorLaser orderrSDen2.5x rSDen

488

50





1



SSC

SSC

488/10

NA


N/AN/A



B530_30B

FITC, GFP, AF488, YFP, 5-FAM, CFSE, BB515, ZsGreen, SYBR Green, BODIPY FL

530/30

505LP


24.761.75


B710_50APerCP-Cy5.5, 7AAD, BB700, PerCP-eFluor710710/50685LP
24.561.25

405

50





2



V440_40FPacific Blue, Cascade Blue, AF405, V450, BV421, Dylight405, DAPI, eFluor450, SparkViolet423, SuperBrigth436440/40NA
20.150.25


V525_50E

CFP, AF430, AmCyan, Cascade Yellow, V500, BV510, PO, eFluor506, SparkViolet538

525/50

505LP

 

23.358.25


V610_20D

Qdot 605, BV605, SuperBright600

610/20

600LP


35.187.75


V660_20C

Qdot 655, BV650, SuperBright645

660/20

630LP


22.656.5


V710_50B

Qdot 700, BV711, SuperBright702

710/50

685LP


28.471


V780_60ABV785, SuperBright780780/60750LP
22.656.5

355

20





3



U379_28CBUV395379/28NA
17.944.75


U515_30BBUV496, DAPI, Hoechst Blue515/30450LP
16.541.25


U740_35ABUV737, Hoechst Red740/35690LP
19.248

640

40





4



R670_14CAPC, AF647, CF647, DiD, DRAQ5, SYTOX RED, eFluor660, SparkNIR685670/14NA
14.235.5


R730_45B

APC-R700, AF700, DRAQ5, FVS700

730/45

690LP


13.734.25


R780_60A

APC-Cy7, APC-H7, DiR, APC-FIRE, APC-eFluor780, APCVio770

780/60

750LP


14.736.75

561

100





5



Y586_15DPE, DsRed, dTomato, mTangerine, mStrawberry, SparkYG581586/15570LP
17.744.25


Y610_20C

PE-Texas Red, mCherry, PE-CF594, AF568, PI, PE-Dazzle, 7AAD, AF594, PE-eFluor610, SparkYG593

610/20600LP
16.441


Y710_50BPE-Cy5.5, 7AAD, PE-Fire700710/50685LP
16.441


Y780_60APE-Cy7, PE-Vio770, 780/60750LP
19.147.75

...

Laser (nm)Power (mw)DetectorCommon fluorochromes detectable (not exhaustive)Band pass filterDichroic MirrorLaser orderrSDen2.5x rSDen

488

50-100

default @ 50





1



SSC

SSC

488/10

N/A


N/AN/A



B530_30B

FITC, GFP, AF488, YFP, 5-FAM, CFSE, BB515, ZsGreen, BODIPY FL, CF488A, DyLight488, EGFP, EYFP, Kiravia Blue520, NovaFluor Blue510, NovaFluor Blue530, SparkBlue550, Sytox Green, VioBright515, VioBright FITC

530/30

505LP


15.137.75


B710_50APerCP-Cy5.5, 7AAD, BB700, PerCP-eFluor710, PE-Fire700, PerCP-Vio700710/50685LP
15.137.75

403

50





2



V440_40FPacific Blue, Cascade Blue, AF405, V450, BV421, Dylight405, DAPI, eFluor450, SparkViolet423, SuperBright436, CellTrace Violet, CF405S, StarBright V440, Sytox Blue, VioBlue440/40N/A
10.125.25


V525_50E

Pacific Green, CFP, AF430, AmCyan, Cascade Yellow, V500, BV510, PO, eFluor506, SparkViolet538, Krome Orange, StarBright V515, T-Sapphire, VioGreen, VioletFluor500

525/50

505LP

 

10.125.25


V610_20D

Qdot 605, BV605, SuperBright600, StarBright V610

610/20

600LP


10.526.25


V660_20C

Qdot 655, BV650, SuperBright645, StarBright V670

660/20

630LP


11.428.5


V710_50B

Qdot 700, BV711, SuperBright702, StarBright V710

710/50

685LP


10.526.25


V780_60ABV785, SuperBright780, StarBright V760, StarBright V790780/60750LP
1025

355

20





3



U379_28CBUV395, StarBright UV400379/28N/A
20.551.25


U515_30BBUV496, DAPI, Hoechst Blue, StarBright UV510515/30450LP
22.155.25


U740_35ABUV737, Hoechst Red, StarBright UV740740_35690LP
20.451

639

40





4



R670_14CAPC, AF647, CF647, DiD, DRAQ5, eFluor660, SparkNIR685, Celltrace Far Red, DyLight633, DyLight649, NovaFluor Red660, NovaFluor Red685, Sytox ORed670/14N/A
15.739.25


R730_45B

APC-R700, AF680, AF700, DRAQ5, FVS700, NovaFluor Red700, R718

730/45

690LP


16.441


R780_60A

APC-Cy7, APC-H7, DiR, APC-Fire750, APC-eFluor780, APCVio770, AF750, APC-AF750

780/60

750LP


1742.5

561

50





5



Y586_15DPE, DsRed, dTomato, mTangerine, mOrange, mStrawberry,SparkYG581, CellTrace Yellow, DyLight549, eFluor570, NovaFluor Yellow570, RY586, Sytox Orange586/15570LP
13.634


Y610_20C

PE-Texas Red, mCherry, PE-CF594, AF568, AF594, PI, PE-Dazzle594, PE eFluor610, SparkYG593, BODPIY 581-591-C11, eFluor615, NovaFluor Yellow610, PE-Vio615, StarBright Y605

610/20600LP
14.937.25


Y710_50BPE-Cy5.5, 7AAD, PE-Fire700, NovaFluor Yellow690, NovaFluor Yellow700, StarBright Y720710/50685LP
13.634


Y780_60APE-Cy7, PE-Vio770, 780/60750LP
15.538.75

...

  • Baseline in use is #23533. 
  • Default voltages are normally good for resolving dim populations. Rule of thumb guide to settings voltages include, ensuring the rSD of the unlabelled cells is at least 2.5x the value of the rSD of electronic noise of the detector (that is viewable from the instrument configuration sheet) that accompanies each cytometer. You need to record a small file in order to accurately determine the rSD of the unlabelled cells each time. Remember this should be the minimum voltage and not the voltage to use. CST voltages are a good voltage to use with the exception that if a population is off scale, then the voltage needs to be reduced.
  • Filter changes on the Fortessa and LSRII can be done once users have been trained. The “Non-default signage” must be displayed when using a non-default filter in place of the default.
  • For optimal detection of PE-CY5, PE-AF647 & mPlum a 670/30 filter is needed in Y710_50B with a 635LP.
  • For optimal detection of PerCP a 675/50 could be used in B710_50A with a 635LP. There are other alternatives too.
  • For optimal detection of Hoechst Blue & AF350 a 450/50 bandpass filter could be used in U515_30B, with a 425LP.
  • For optimal detection of AF633 a 670/40 bandpass filter could be used in R670_14C.

Symphony

Baseline:  

Baseline report: 

Cytometer Baseline Reportsymphony.pdf

...

Laser ColourLaser (nm)Max power (mw)CST default power (mw)DetectorCommon fluorochromes detectable (not exhaustive)Band pass filterDichroic mirrorNotch filterLaser orderPinhole positionDetector specificsOrientationAdditional notes
Red637140140




11
5D




R820_60APC-Cy7, APC-H7, DiR, APC-Vio770, APC-Fire810820/60 BP800LP


H14121-50





R780_60AF750, APC-AF750, APC-eFluor780, APC-Fire750, APC-H7780/60 BP750 LP785nm

H14121-50





R730_45APC-R700, AF700, DRAQ5, FVS700, AF680, NovaFluor Red685, NovaFluor Red700, R718730/45 BP685 LP


H11900-20





R670_30APC, AF647, CF647, DiD, DRAQ5, eFluor660, Spark NIR685, CellTrace Far Red, DyLight633, DyLight649, NovaFluor Red660, Sytox Red670/30 BP665 LP


H11900-20
Schott RG665
UV355100100




23
M1&M2&M1




U820_60BUV805820/60 BP800 LP785nm

H14121-50





U780_60spectral780/60 BP770 LP


H14121-50





U740_35BUV737, Hoechst Red, StarBright UV740740/35 BP710 LP


H14121-50





U695_40
695/40685 LP









U670_30BUV661, StarBright UV665670/30 BP635 LP


H11900-20





U610_20BUV615, StarBright UV610, StarBright UV605610/20 BP600 LP


H11900-20





U586_15BUV563586/15 BP570 LP594nm

H11900-20





U515_30BUV496, DAPI, Hoechst Blue, StarBright UV510515/30 BP450 LP


H11900-20





U450_50AF350, DAPI, CellTrace Blue, StarBright UV445450/50 BP410 LP


H11900-20





U379_28BUV395, StarBright UV400379/28 BP



H11900-110

Yellow561150150




34
6D




Y820_60PE-Cy7, StarBright Y800820/60 BP800 LP


H14121-50





Y780_60
780/60 BP750 LP785nm

H11900-20





Y710_50PE-Cy5.5, PE-Fire700, NovaFluor Yellow690, NovaFluor Yellow700, StarBright Y720710/50 BP685 LP


H11900-20





Y670_30PE-Cy5, NovaFluor Yellow660, StarBright Y665670/30 BP635 LP


H11900-20





Y610_20PE-CF594, PE-Texas Red, mCherry, AF568, AF594, PI, PE-Dazzle594, PE eFluor610, SparkYG593, NovaFluor Yellow610, PE-Vio615, StarBright Y605610/20 BP600 LP


H11900-20





Y586_15PE, DsRed, dTomato, mTangerine, mOrange, mStrawberry, Spark YG581, CellTrace Yellow, DyLight549, eFluor570, NovaFluor Yellow570, R586, Sytox Orange586/15 BP

594nm

Y-58 Schott



H11900-40

Blue488200150




45
M1&M2




B820_60BB790, StarBright Blue810820/60 BP800 LP


H14121-50





B780_60RB780780/60 BP750 LP785nm

H14121-50





B750_30BB755, StarBright Blue 765750/30 BP735 LP


H14121-50





B710_50BB700, PerCP-Cy5.5, 7AAD, PerCP-eFluor710, PerCP-Vio700, PE-Fire700, StarBright Blue700710/50 BP685 LP


H11900-20





B670_30BB660, PerCP, NovaFluor Blue660-40S, NovaFluor Blue610-120S, StarBright Blue675670/30 BP635 LP


H11900-20





B610_20BB630, NovaFluor Blue610-30S, NovaFluor Blue610-70S, StarBright Blue615610/20 BP600 LP594nm

H11900-20





B515_30FITC, BB515, A488, BODIPY FL, CF488A, CSFE, DyLight488, EGFP, Kiravia Blue510, NF-Blue510, NF-Blue530, Spark Blue550, Sytox Green, VioBright515, VioBright FITC515/30 BP500 LP


H11900-20





SSC
488/10 BP



H11900-20

Orange594100100




56
5D




O660_20spectral660/20 BP635 LP


H14121-20





O616_23AF594, mCherry616/23 BP610 LP


H14121-20

Violet406200200




67
M1&M2&M1




V820_60BV786, BV875, SuperBright780, StarBright V790820/60 BP800 LP


H14121-50





V780_60spectral780/60 BP770 LP785nm 

H14121-50





V750_30BV750, StarBright V760750/30 BP735 LP


H14121-50





V710_50BV711, Qdot700, SuperBright702, StarBright V710710/50 BP685 LP


H11900-20





V677_20BV650, Qdot650, SuperBright645, StarBright V670677/20 BP635 LP


H11900-20





V610_20BV605, Qdot605, SuperBright600, StarBright V610610/20 BP600 LP


H11900-20





V586_15BV570, Pacific Orange586/15 BP550 LP594nm

H11900-20





V525_50BV510, Pacific Green, CFP, AF430, AmCyan, Cascade Yellow, V500, PO, eFluor506, Spark Violet538, StarBright V515, T-Sapphire, VioGreen, VioletFluor500525/50 BP505 LP


H11900-20





V474_25BV480, DAPI, StarBright V475474/25 BP450 LP


H11900-20





V427_25BV421, Pacific Blue, Cascade Blue, AF405, V450, eFluor450, VioBlue,  Spark Violet423, StarBright V440, SuperBright436, Sytox Blue427/25 BP410 LP


H11900-20





V405_10Violet SSC405/10 BP



H11900-20







*PMT installed



H11900-20







*PMT installed



H11900-20







*PMT installed



H11900-20

IndigoNIR446_77975_10075_100




79
5D




N820_60DyLight800 (strep-avidin - recommended), AF790820/60 BP800 LP


H14121-50





I660_20spectral660/20 BP635 LP


H14121-20





I605_40spectral605/40 BP600 LP


H11900-20





I515_30spectral515/30 BP500 LP


H11900-20





I470_15CFP, Chromomycin470/15 BP450 LP


H11900-20

...

Laser (nm)

Power

(mw)

DetectorCommon fluorochromes detectable (not exhaustive)Band pass filterDichroic MirrorLaser order

488

50-200

default 100





1



PMT 1 - FSC (Par)Forward Scatter - Parrallel488/10NA



PMT 2 - FSC (Perp)Forward Scatter - Perpendicular488/10NA


PMT 3 - SSCSide Scatter488/10NA


PMT 4FITC, GFP, AF488, BBYFP, 5-FAM, CFSE , BB515, ZsGreen, DiO, BODIPY FL530/30600LP


PMT 5PerCP, PerCP-Cy5.5, 7AAD, PI, PerCP-eFL710, PerCP-Vio700, BB700, PerCP-eFluor710710/50NA
405100405 nm - 100mw   2


PMT 14Pacific Blue, Cascade Blue, AF405, V450, BV421, DyLight 405, eFL450, VioBlue, DAPI, eFluor450440/40455LP, 500LP


PMT 15CFP, eCFP, eFluor506480/40500LP


PMT 16CFP, AF430, AmCyan, Cascade Yellow, V500, BV510, PO, VioGreen525/30550LP


PMT 17BV570, Qdot 565, 572/28585LP, 645LP


PMT 18BV605, DyLight405, Qdot 605, SuperBright600610/20645LP


PMT 19BV650, eFL650, Qdot 655, SuperBright645660/20685LP


PMT 20BV711, Qdot 700, SuperBright702710/50NA
561150561 nm - 150mw   3


PMT 6PE, DsRed, dTomato, mTangerine, mStrawberry, Dil585/29600LP


PMT 7PE-Texas Red, mCherry, PE-CF594, PE-AF610, AF568, PE-eFL610, PE-Dazzle594,  PE eFluor610610/20645LP


PMT 8PE-Cy5670/30685LP


PMT 9PE-Cy5.5710/50740LP


PMT 10PE-Cy7, PE-Vio770750LPNA
355100355 nm - 100mw   4


PMT 13DAPI, Hoechst Blue460/50480LP, 600LP


PMT 22BUV395379/34427LP


PMT 23BUV496520/35550LP, 600LP


PMT 24BUV661, Hoechst Red670/30NA


PMT_UnpluggedBUV563585/42NA
628120640 nm - 120mw   5


PMT 11APC, AF647, CF647, DiD, DRAQ5,eFluor660670/30700LP


PMT 12AF700, DRAQ5, APC-R700740/40740LP


PMT 21APC-Cy7, APC-H7, DiR, APC-eFL780, APC-Fire750750LPNA

...

Laser (nm)

Power (mw)DetectorCommon fluorochromes detectable (not exhaustive)Band pass filterDichroic MirrorLaser order

405

50





2


V405_10

vSSC

405/10

NA




V450_50

DAPI, BV421, Pacific Blue, Cascade Blue, AF405, V450, DyLight 405, eFL450, VioBlue

450/45

NA


488 

20





1


SSC

SSC

488/8

NA




B525_40

FITC, GFP, AF488, YFP, 5-FAM, CFSE, BB515, ZsGreen, SYBR Green

525/40

NA




B585_42

PE,PI,DsRed,DiA

585/42

NA


633

20

NOT ACTIVENOT ACTIVE

3

...